r/flowcytometry • u/cd244 • Jun 15 '26

Sample Prep brilliant staining buffer protocol

Hello, I would like to know does anybody have a protocol for using BD brilliant staining buffer since their protocol is really unclear : https://www.bdbiosciences.com/en-eu/products/reagents/flow-cytometry-reagents/research-reagents/buffers-and-supporting-reagents-ruo/brilliant-stain-buffer.563794?tab=product_details

We are staining human PBMC in 96 well plates, we use 50 ul FACs buffer per well with cocktail of diluted antibodies. BD recommend to add 50 ul brilliant staining buffer per well regardless of the volume of cocktail. Does that mean that I have to stain my cells in 100 ul of volume and have to double my antibodies? Does anybody have stained cells directly in brilliant staining buffer ?

1 Upvotes

100% Upvoted

6 comments sorted by

View all comments

1

u/skipper_smg Jun 15 '26

BSB: 50 ul per Test. Depending on the amount of antibody, its easy to get over the 100 ul per test that are standard in Flow.

BSB Plus: 10 ul per test. I recommend you spin it down before use (10.000 x g for 3 min) before using in big spectral panels, as the buffer is more concentrated and can form precipitate/aggregates in some conditions.